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https://www.readbyqxmd.com/read/28729350/structures-of-the-crispr-genome-integration-complex
#1
Addison V Wright, Jun-Jie Liu, Gavin J Knott, Kevin W Doxzen, Eva Nogales, Jennifer A Doudna
CRISPR-Cas systems depend on the Cas1-Cas2 integrase to capture and integrate short foreign DNA fragments into the CRISPR locus, enabling adaptation to new viruses. We present crystal structures of Cas1-Cas2 bound to both donor and target DNA in intermediate and product integration complexes, as well as a cryo-electron microscopy structure of the full CRISPR locus integration complex including the accessory protein Integration Host Factor (IHF). The structures show unexpectedly that indirect sequence recognition dictates integration site selection by favoring deformation of the repeat and the flanking sequences...
July 20, 2017: Science
https://www.readbyqxmd.com/read/28728288/-descemet-s-membrane-endothelial-keratoplasty-for-treatment-of-patients-with-corneal-endothelial-decompensation
#2
T Huang, C Ouyang, J Zhan, L Jiang
Objective: To evaluate the clinical outcomes of Descemet's membrane endothelial keratoplasty (DMEK) for treatment of patients with corneal endothelial decompensation in China. Methods: Interventional case series. Ten patients (10 eyes) with the diagnosis of corneal endothelial decompensation received DMEK during July 2016 and September 2016 in Zhongshan Ophthalmic Center. Descemet's membrane of the donor tissue was carefully stripped from the underlying stroma manually and cut with a 7.75- or 8.0-mm-diameter donor punch...
July 11, 2017: [Zhonghua Yan Ke za Zhi] Chinese Journal of Ophthalmology
https://www.readbyqxmd.com/read/28724956/mapping-resistance-responses-to-sclerotinia-infestation-in-introgression-lines-of-brassica-juncea-carrying-genomic-segments-from-wild-brassicaceae-b-fruticulosa
#3
Kusum Rana, Chhaya Atri, Mehak Gupta, Javed Akhatar, Prabhjodh S Sandhu, Nitin Kumar, Ravinder Jaswal, Martin J Barbetti, Surinder S Banga
Sclerotinia stem rot (Sclerotinia sclerotiorum) is a major disease of Brassica oilseeds. As suitable donors to develop resistant cultivars are not available in crop Brassicas, we introgressed resistance from a wild Brassicaceae species, B. fruticulosa. We produced 206 B. juncea-B. fruticulosa introgression lines (ILs). These were assessed for pollen grain fertility, genome size variations and resistance responses to Sclerotinia following stem inoculations under disease-conducive conditions. Of these, 115 ILs showing normal fertility and genome size were selected for cytogenetic characterization using florescent genomic in situ hybridization (Fl-GISH)...
July 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28714989/correction-of-a-splicing-defect-in-a-mouse-model-of-congenital-muscular-dystrophy-type-1a-using-a-homology-directed-repair-independent-mechanism
#4
Dwi U Kemaladewi, Eleonora Maino, Elzbieta Hyatt, Huayun Hou, Maylynn Ding, Kara M Place, Xinyi Zhu, Prabhpreet Bassi, Zahra Baghestani, Amit G Deshwar, Daniele Merico, Hui Y Xiong, Brendan J Frey, Michael D Wilson, Evgueni A Ivakine, Ronald D Cohn
Splice-site defects account for about 10% of pathogenic mutations that cause Mendelian diseases. Prevalence is higher in neuromuscular disorders (NMDs), owing to the unusually large size and multi-exonic nature of genes encoding muscle structural proteins. Therapeutic genome editing to correct disease-causing splice-site mutations has been accomplished only through the homology-directed repair pathway, which is extremely inefficient in postmitotic tissues such as skeletal muscle. Here we describe a strategy using nonhomologous end-joining (NHEJ) to correct a pathogenic splice-site mutation...
July 17, 2017: Nature Medicine
https://www.readbyqxmd.com/read/28711250/genetic-analysis-for-quality-of-frozen-embryos-produced-by-holstein-cattle-donors-in-canada
#5
C Jaton, F S Schenkel, F Malchiodi, M Sargolzaei, C A Price, C Baes, F Miglior
The number of embryos produced by Holstein donors has been shown to be heritable, so it could be possible to genetically select for this trait to improve the efficiency of the assisted reproductive technology (ART) in dairy cattle. Another important parameter to consider for achieving good results from ART is embryo quality because embryos of good quality have more chance of producing live offspring. The possibility of using genetic selection for increasing the quality of embryo produced from ART has yet to be assessed...
July 12, 2017: Journal of Dairy Science
https://www.readbyqxmd.com/read/28709845/fifteen-years-of-nucleic-acid-testing-in-france-results-and-lessons
#6
S Laperche, P Tiberghien, C Roche-Longin, J Pillonel
Of the 40 million donations screened with Nucleic acid testing (NAT) between July 2001 and December 2015 in France, 20 HIV-positive, 13 HCV-positive and 17 HBV (HBV-NAT was initiated in 2005 and extended to the whole country in 2010) donations were discarded thanks to NAT. The main benefit in terms of discarded donations is related to HBV with a yield of 0.88 per million donations, which is 12.5 and 1.8 times higher than for HCV and HIV respectively. The main risk factor found in these donors during the post donation interview was having sex with men for males (n=11, all repeat blood donors), having a partner HCV positive (n=6) or at-risk partner (originated from endemic area or HBV positive) for HBV (n=8) for HIV, HCV and HBV, respectively...
July 11, 2017: Transfusion Clinique et Biologique: Journal de la Société Française de Transfusion Sanguine
https://www.readbyqxmd.com/read/28709085/amplification-of-electrochemical-signal-by-a-whole-cell-redox-reactivation-module-for-ultrasensitive-detection-of-pyocyanin
#7
Yuan Yang, Yang-Yang Yu, Yan-Zhai Wang, Chun-Lian Zhang, Jing-Xian Wang, Zhen Fang, Huoyang Lv, Jian-Jiang Zhong, Yang-Chun Yong
A bioelectrochemical sensing system based on a novel whole-cell redox reactivation/cycling module for ultrasensitive detection of pyocyanin (the biomarker of Pseudomonas aeruginosa infections) was developed. The electroactive bacteria mediated redox reactivation module was constructed using Shewanella oneidensis MR-1 cells as the bioelectro-catalyst and lactate as the electron donor. It could regenerate reductive pyocyanin from its oxidative state, which enabled pyocyanin molecule repeatedly registered by the electrode...
July 5, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28694465/generation-of-complement-protein-c3-deficient-pigs-by-crispr-cas9-mediated-gene-targeting
#8
Wei Zhang, Guan Wang, Ying Wang, Yong Jin, Lihua Zhao, Qiang Xiong, Lining Zhang, Lisha Mou, Rongfeng Li, Haiyuan Yang, Yifan Dai
Complement protein C3 is the pivotal component of the complement system. Previous studies have demonstrated that C3 has implications in various human diseases and exerts profound functions under certain conditions. However, the delineation of pathological and physiological roles of C3 has been hampered by the insufficiency of suitable animal models. In the present study, we applied the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system to target the C3 gene in porcine fetal fibroblasts...
July 10, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28692815/efficient-generation-of-goats-with-defined-point-mutation-i397v-in-gdf9-through-crispr-cas9
#9
Yiyuan Niu, Xiaoe Zhao, Jiankui Zhou, Yan Li, Yu Huang, Bei Cai, Yutai Liu, Qiang Ding, Shiwei Zhou, Jin Zhao, Guangxian Zhou, Baohua Ma, Xingxu Huang, Xiaolong Wang, Yulin Chen
The recent emergence of the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) 9 system has attracted significant attention for its potential to improve traits of agricultural importance. However, most applications in livestock species to date have depended on aberrant DNA repair to generate frameshifting indels. Whether this genomic engineering technique involving homology-dependent repair (HDR) can be used to introduce defined point mutations has been less explored. Previously, we reported a G→A point mutation (g...
July 11, 2017: Reproduction, Fertility, and Development
https://www.readbyqxmd.com/read/28690606/a-highly-efficient-crispr-cas9-mediated-large-genomic-deletion-in-bacillus-subtilis
#10
Younju So, Soo-Young Park, Eun-Hye Park, Seung-Hwan Park, Eui-Joong Kim, Jae-Gu Pan, Soo-Keun Choi
In Bacillus subtilis, large genomic deletions have been carried out for genome reduction, antibiotic overproduction, and heterologous protein overexpression. In view of the eco-friendliness of B. subtilis, it is critical that engineering preserves its food-grade status and avoids leaving foreign DNA in the genome. Existing methods of generating large genomic deletions leave antibiotic resistance markers or display low mutation efficiency. In this study, we introduced a clustered regularly interspaced short palindromic repeat-derived genome engineering technique to develop a highly efficient method of generating large genomic deletions in B...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28685174/amine-functionalized-mil-53-al-coated-stainless-steel-fiber-for-efficient-solid-phase-microextraction-of-synthetic-musks-and-organochlorine-pesticides-in-water-samples
#11
Lijun Xie, Shuqin Liu, Zhubing Han, Ruifen Jiang, Fang Zhu, Weiqin Xu, Chengyong Su, Gangfeng Ouyang
The fiber coating is the key part of the solid-phase microextraction (SPME) technique, and it determines the sensitivity, selectivity, and repeatability of the analytical method. In this work, amine (NH2)-functionalized material of Institute Lavoisier (MIL)-53(Al) nanoparticles were successfully synthesized, characterized, and applied as the SPME fiber coating for efficient sample pretreatment owing to their unique structures and excellent adsorption properties. Under optimized conditions, the NH2-MIL-53(Al)-coated fiber showed good precision, low limits of detection (LODs) [0...
July 6, 2017: Analytical and Bioanalytical Chemistry
https://www.readbyqxmd.com/read/28683668/kinship-identities-in-the-context-of-uk-maternal-spindle-transfer-and-pronuclear-transfer-legislation
#12
Calum MacKellar
In the discussions leading up to the enactment of the UK Human Fertilisation and Embryology (Mitochondrial Donation) Regulations 2015, it was repeatedly emphasised, by many commentators, that maternal spindle transfer (MST) and pronuclear transfer (PNT) did not give rise to children who could be considered as having three or more parents. This was because it was argued that only the genetic material found in the chromosomes should be considered as the determining factor for the formation of parent-child relationships and the resulting kinship identities...
July 6, 2017: New Bioethics: a Multidisciplinary Journal of Biotechnology and the Body
https://www.readbyqxmd.com/read/28682619/structure-reactivity-relationships-of-conformationally-armed-disaccharide-donors-and-their-use-in-the-synthesis-of-a-hexasaccharide-related-to-the-capsular-polysaccharide-from-streptococcus-pneumoniae-type-37
#13
Thibault Angles d'Ortoli, Christoffer Hamark, Göran Widmalm
To advance the field of glycobiology, efficient synthesis methods of oligosaccharides and glycoconjugates are a requisite. In glycosylation reactions using superarmed donors, both selectivity and reactivity issues must be considered, and we herein investigate these aspects for differently protected β-linked 2-O-glycosylated glucosyl donors carrying bulky tert-butyldimethylsilyl groups to different extents. The acceptors in reactions being secondary alcohols presents a challenging situation with respect to steric crowding...
July 19, 2017: Journal of Organic Chemistry
https://www.readbyqxmd.com/read/28671277/a-motivational-interview-promotes-retention-of-blood-donors-with-high-internal-motivation
#14
Christopher R France, Janis L France, Bruce W Carlson, Lina K Himawan, Debra A Kessler, Mark Rebosa, Beth H Shaz, Katrala Madden, Patricia M Carey, P Maxwell Slepian, Brett Ankawi, Irina E Livitz, Kristen R Fox
BACKGROUND: Based on the hypothesis that self-determined motivation is associated with an increased likelihood of future behavior, the present study examined the ability of a motivational interview to promote internal motivation for giving blood and future donation attempts. STUDY DESIGN AND METHODS: A sample of 484 recent whole-blood and double red blood cell donors (62.4% female; age = 30.2 ± 11.8 years) were randomly assigned to either a telephone-delivered motivational interview or a control call approximately 6 weeks after donating...
July 3, 2017: Transfusion
https://www.readbyqxmd.com/read/28665587/organohalide-respiration-with-chlorinated-ethenes-under-low-ph-conditions
#15
Yi Yang, Natalie L Cápiro, Tyler F Marcet, Jun Yan, Kurt D Pennell, Frank E Löffler
Bioremediation at chlorinated solvent sites often leads to groundwater acidification due to electron donor fermentation and enhanced dechlorination activity. The microbial reductive dechlorination process is robust at circumneutral pH, but activity declines at groundwater pH values below pH 6.0. Consistent with this observation, the activity of tetrachloroethene (PCE) dechlorinating cultures declined at pH 6.0 and was not sustained in pH 5.5 medium, with one notable exception. Sulfurospirillum multivorans dechlorinated PCE to cis-1,2-dichloroethene (cDCE) in pH 5...
June 30, 2017: Environmental Science & Technology
https://www.readbyqxmd.com/read/28662917/accumulative-scfv-fc-antibody-gene-integration-into-the-hprt-chromosomal-locus-of-chinese-hamster-ovary-cells
#16
Xue Wang, Yoshinori Kawabe, Risa Kato, Takeshi Hada, Akira Ito, Yoshimasa Yamana, Masako Kondo, Masamichi Kamihira
We have previously developed an accumulative site-specific gene integration system (AGIS) using Cre-recombinase and mutated loxP sites. AGIS enables repeated transgene integration into a predetermined chromosomal site in mammalian cells. However, the process of establishing cells with multiple integrated copies of the transgene is still time-consuming. In the present study, we describe an improved version of AGIS that facilitates and accelerates the establishment of high-producer Chinese hamster ovary (CHO) cells...
June 26, 2017: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28660316/expression-of-flp-protein-in-a-baculovirus-insect-cell-system-for-biotechnological-applications
#17
Ida S Jensen, Ken Inui, Srdja Drakulic, Sakthidasan Jayaprakash, Bjoern Sander, Monika M Golas
The Saccharomyces cerevisiae Flp protein is a site-specific recombinase that recognizes and binds to the Flp recognition target (FRT) site, a specific sequence comprised of at least two inverted repeats separated by a spacer. Binding of four monomers of Flp is required to mediate recombination between two FRT sites. Because of its site-specific cleavage characteristics, Flp has been established as a genome engineering tool. Amongst others, Flp is used to direct insertion of genes of interest into eukaryotic cells based on single and double FRT sites...
August 2017: Protein Journal
https://www.readbyqxmd.com/read/28659431/odor-identification-as-a-biomarker-of-preclinical-ad-in-older-adults-at-risk
#18
Marie-Elyse Lafaille-Magnan, Judes Poirier, Pierre Etienne, Jennifer Tremblay-Mercier, Joanne Frenette, Pedro Rosa-Neto, John C S Breitner
OBJECTIVE: To assess odor identification (OI) as an indicator of presymptomatic Alzheimer disease (AD) pathogenesis in cognitively normal aging individuals at increased risk of AD dementia. METHODS: In 274 members of the PREVENT-AD cohort of healthy aging persons with a parental or multiple-sibling history of AD dementia, we assessed the cross-sectional association of OI with potential indicators of presymptomatic AD. Some 101 participants donated CSF, thus enabling assessment of AD pathology with the biomarkers total tau (t-tau), phospho-tau (P181-tau), and their ratios with β-amyloid (Aβ1-42)...
June 28, 2017: Neurology
https://www.readbyqxmd.com/read/28653765/a-liquid-calcium-vitamin-d3-supplement-is-effective-prophylaxis-against-hypocalcemic-toxicity-during-apheresis-platelet-donation
#19
Robert Weinstein, Stefanie Haynes, Yong Zhao, Elda Hickson, Jeanne Linden, Patricia St Pierre, Paula Ducharme, Paula Sulmasy, Molly Graves, Jeffrey A Bailey, Linda Welch, Amie Simard, Michelle Vauthrin, Mindy Greene
Hypocalcemic toxicity, because of return of citrate anion to the donor, is the major toxicity of apheresis platelet donation. Oral calcium carbonate, given prophylactically at the start of donation, has shown limited ability to alleviate this toxicity. We examined whether repeated prophylactic doses of calcium carbonate, or of a liquid preparation containing calcium citrate, calcium phosphate, and vitamin D3 , would be more effective at preventing symptoms of hypocalcemic toxicity. Symptoms were reported by 48% of donors who received no prophylaxis and 60% of donors who received 1000 mg of oral calcium carbonate at the start of, and every 20 minutes during, donation (P = 0...
June 27, 2017: Journal of Clinical Apheresis
https://www.readbyqxmd.com/read/28649744/mannose-binding-lectin-deficient-donors-increase-the-risk-of-bacterial-infection-and-bacterial-infection-related-mortality-after-liver-transplantation
#20
J Lombardo, G Sanclemente, J Colmenero, M Español-Rego, M T Arias, P Ruiz, E Mauro, L Sastre, G Crespo, A Rimola, A Moreno, F Lozano, M Navasa
Mannose-binding lectin (MBL) is synthesized by the liver and binds to microbes. MBL2 gene polymorphisms produce intermediate/low/null- or normal-MBL serum levels (MBL-deficient or MBL-sufficient phenotypes, respectively). We aimed to evaluate the incidence and severity of infection, rejection and survival within 1 year after liver transplantation (LT) according to donor and recipient MBL2 gene polymorphisms. A repeated-event analysis for infection episodes (negative binomial regression, Andersen-Gill model) was performed in 240 LT...
June 26, 2017: American Journal of Transplantation
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